Measurement of circulating cell-derived microparticles by flow cytometry: Sources of variability within the assay

Glycoproteins of GpIbα and GpIIbIIIa on the Synthetic or Naturally Occurred Platelet-Derived Microparticles

“…For example nPMP can bind fibrinogen and participate in thrombus formation [13]. In this research, the retention of gpIIb/IIIa and gpIba molecules was demonstrated on the surface of plateletderived IPM microparticles using flow cytometry that is the usual method for the study of microparticles [18]. The obtained results had conflict with the previous report related to the absence of gpIIb/IIIa molecules on the surface of produced IPM [10].…”

Does RBC Storage Age Effect Inflammation, Immune Function and Susceptibility to Transfusion Associated Microchimerism in Critically Ill Patients? Adverse Effects of RBC Storage in Critically Ill Patients

“…Of note, longer centrifugation times than we used in the current experiments have been described (69), but we only tested a 5 min centrifugation in the current work in an attempt to develop a protocol suited to high throughput sam ple analysis. Similar 5 min centrifugation protocols to remove antibody aggregates were recommended by other researchers as well, including 16,000g for 5 min (70) and 18,000g for 5 min (63). Antibodies filtered with smaller pore sizes were equally as effective as those filtered with larger pore sizes.…”

“…EV sample collection and processing is yet another area in which standardization is needed, yet no con sensus exists on an optimal protocol (59 61). Many different pre analytical variables have been shown to affect EV content, including storage temperature and duration (62,63), anticoa gulant/preservative used (62,64), and centrifugation method used (59,63). Specialized techniques and optimized protocols have been recommended (61); however, there is no consensus on the best method for EV detection by FCM.…”

Microvesicles and diabetic complications — novel mediators, potential biomarkers and therapeutic targets

“…Increasing the time between venepuncture and centrifugation and a single freeze-thaw cycle of samples lead to increased levels of MVs [31] . Washing samples, double centrifugation of MVs prior to freezing, and long-term storage of MV samples at -80°C result in decreased MV blood levels [31] .…”

“…Increasing the time between venepuncture and centrifugation and a single freeze-thaw cycle of samples lead to increased levels of MVs [31] . Washing samples, double centrifugation of MVs prior to freezing, and long-term storage of MV samples at -80°C result in decreased MV blood levels [31] . The three major parameters are the delay before the first centrifugation, agitation of the tubes during transportation and the centrifugation protocol [27] .…”